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Cell DOI:10.1016/j.cell.2015.06.059

A Genome-wide CRISPR Screen in Primary Immune Cells to Dissect Regulatory Networks.

Publication TypeJournal Article
Year of Publication2015
AuthorsParnas, O, Jovanovic, M, Eisenhaure, TM, Herbst, RH, Dixit, A, Ye, CJimmie, Przybylski, D, Platt, RJ, Tirosh, I, Sanjana, NE, Shalem, O, Satija, R, Raychowdhury, R, Mertins, P, Carr, SA, Zhang, F, Hacohen, N, Regev, A
Date Published2015 Jul 30
KeywordsAnimals, Bone Marrow Cells, Cell Differentiation, Cell Survival, CRISPR-Cas Systems, Dendritic Cells, Gene Knockout Techniques, Gene Regulatory Networks, Genetic Techniques, Hexosyltransferases, Immunity, Innate, Membrane Proteins, Mice, Mice, Transgenic, Toll-Like Receptor 4, Tumor Necrosis Factor-alpha

Finding the components of cellular circuits and determining their functions systematically remains a major challenge in mammalian cells. Here, we introduced genome-wide pooled CRISPR-Cas9 libraries into dendritic cells (DCs) to identify genes that control the induction of tumor necrosis factor (Tnf) by bacterial lipopolysaccharide (LPS), a key process in the host response to pathogens, mediated by the Tlr4 pathway. We found many of the known regulators of Tlr4 signaling, as well as dozens of previously unknown candidates that we validated. By measuring protein markers and mRNA profiles in DCs that are deficient in known or candidate genes, we classified the genes into three functional modules with distinct effects on the canonical responses to LPS and highlighted functions for the PAF complex and oligosaccharyltransferase (OST) complex. Our findings uncover new facets of innate immune circuits in primary cells and provide a genetic approach for dissection of mammalian cell circuits.


Alternate JournalCell
PubMed ID26189680
PubMed Central IDPMC4522370
Grant ListR01 MH110049 / MH / NIMH NIH HHS / United States
P30 DK043351 / DK / NIDDK NIH HHS / United States
K99 HG008171 / HG / NHGRI NIH HHS / United States
5DP1-MH100706 / DP / NCCDPHP CDC HHS / United States
K99-HG008171 / HG / NHGRI NIH HHS / United States
/ / Howard Hughes Medical Institute / United States
DP1 MH100706 / MH / NIMH NIH HHS / United States
P50 HG006193 / HG / NHGRI NIH HHS / United States
R01 DK097768 / DK / NIDDK NIH HHS / United States
5R01-DK097768 / DK / NIDDK NIH HHS / United States